Kaposi's Sarcoma-Associated Herpesvirus K8 Is an RNA Binding Protein That Regulates Viral DNA Replication in Coordination with a Noncoding RNA
Kaposi's Sarcoma-Associated Herpesvirus K8 Is an RNA Binding Protein That Regulates Viral DNA Replication in Coordination with a Noncoding RNA
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卡波西肉瘤相关疱疹病毒 K8 是一种 RNA 结合蛋白，可与非编码 RNA 协调调节病毒 DNA 复制
DOI：
10.1128/jvi.02177-17
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发表时间：
2018-01
影响因子：
5.4
通讯作者：
Yan Yuan
中科院分区：
文献类型：
--
作者：
Dongcheng Liu;Yan Wang;Yan Yuan
卡波济肉瘤相关疱疹病毒（KSHV）的裂解性复制和新鲜细胞的持续初次感染对病毒致瘤性至关重要。病毒编码的bZIP家族蛋白K8在病毒再活化和从头感染中的病毒DNA复制中起重要作用。K8在病毒生命周期中的功能作用的机制是难以捉摸的。在这里，我们报告说，K8是一种RNA结合蛋白，也与许多其他蛋白质，包括其他RNA结合蛋白。许多涉及K8的蛋白质-蛋白质相互作用由RNA介导。使用结合高通量测序的UV交联和免疫沉淀（CLIP）程序，鉴定了BCBL-1细胞中与K8相关的RNA，包括病毒（PAN、T1.4、T0.7等）和RNA。和蜂窝（MALAT-1、MRP、7SK等）RNA。在K8中定义了一个RNA结合基序，该基序的突变消除了K8与许多非编码RNA结合的能力，以及从头感染期间病毒DNA复制的能力，这表明K8在病毒复制中的功能是通过RNA缔合来进行的。详细研究了K8和相关T1.4 RNA的功能，结果表明T1.4介导K8与ori-Lyt DNA的结合。T1.4-K8复合物与KSHV ori-Lyt DNA物理结合，并招募其他蛋白质和辅因子组装成复制复合物。T1.4的缺失消除了原发感染中的DNA复制。这些发现为K8与T1.4 RNA协调调节KSHV从头感染期间DNA复制的作用提供了机制见解。重要性哺乳动物转录组的全基因组分析显示，以前注释为非编码区的大部分序列实际上被转录并产生稳定的RNA。大量非编码RNA的出现表明功能性RNA-蛋白质复合物，例如，核糖体或剪接体不是最后一个核糖有机体的古老遗迹，但将很好地适应生物学中的调节作用。K8因其独特的特性而一直令人困惑，例如在基因表达和DNA复制中的多重调节作用，而没有DNA结合能力。这项研究揭示了其调节作用的机制，证明K8是一种RNA结合蛋白，与DNA结合，并与非编码RNA协同启动DNA复制。这表明，许多K8功能，如果不是全部，是通过其相关的RNA进行。
ABSTRACT Kaposi's sarcoma-associated herpesvirus (KSHV) lytic replication and constant primary infection of fresh cells are crucial for viral tumorigenicity. The virus-encoded bZIP family protein K8 plays an important role in viral DNA replication in both viral reactivation and de novo infection. The mechanism underlying the functional role of K8 in the viral life cycle is elusive. Here, we report that K8 is an RNA binding protein that also associates with many other proteins, including other RNA binding proteins. Many protein-protein interactions involving K8 are mediated by RNA. Using a UV cross-linking and immunoprecipitation (CLIP) procedure combined with high-throughput sequencing, RNAs that are associated with K8 in BCBL-1 cells were identified, including both viral (PAN, T1.4, T0.7, etc.) and cellular (MALAT-1, MRP, 7SK, etc.) RNAs. An RNA binding motif in K8 was defined, and mutation of the motif abolished the ability of K8 to bind to many noncoding RNAs, as well as viral DNA replication during de novo infection, suggesting that the K8 functions in viral replication are carried out through RNA association. The functions of K8 and associated T1.4 RNA were investigated in detail, and the results showed that T1.4 mediates the binding of K8 to ori-Lyt DNA. The T1.4-K8 complex physically bound to KSHV ori-Lyt DNA and recruited other proteins and cofactors to assemble a replication complex. Depletion of T1.4 abolished DNA replication in primary infection. These findings provide mechanistic insights into the role of K8 in coordination with T1.4 RNA in regulating KSHV DNA replication during de novo infection. IMPORTANCE Genomewide analyses of the mammalian transcriptome revealed that a large proportion of sequence previously annotated as noncoding regions is actually transcribed and gives rise to stable RNAs. The emergence of a large number of noncoding RNAs suggests that functional RNA-protein complexes, e.g., ribosomes or spliceosomes, are not ancient relics of the last ribo-organism but would be well adapted to a regulatory role in biology. K8 has been puzzling because of its unique characteristics, such as multiple regulatory roles in gene expression and DNA replication without DNA binding capability. This study reveals the mechanism underlying its regulatory role by demonstrating that K8 is an RNA binding protein that binds to DNA and initiates DNA replication in coordination with a noncoding RNA. It is suggested that many K8 functions, if not all, are carried out through its associated RNAs.
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影响因子：
8.8
作者：
Weyn-Vanhentenryck SM;Mele A;Yan Q;Sun S;Farny N;Zhang Z;Xue C;Herre M;Silver PA;Zhang MQ;Krainer AR;Darnell RB;Zhang C
通讯作者：
Zhang C
影响因子：
7.3
作者：
Peterlin, B. Matija;Brogie, John E.;Price, David H.
通讯作者：
Price, David H.
影响因子：
5.4
作者：
Yan Wang;Hong Li;Man Yee Chan;F. Zhu;D. Lukac;Yan Yuan
通讯作者：
Yan Wang;Hong Li;Man Yee Chan;F. Zhu;D. Lukac;Yan Yuan
DOI：
10.3390/v6114212
发表时间：
2014-11-04
期刊：
Viruses
影响因子：
--
作者：
Rossetto CC;Pari GS
通讯作者：
Pari GS
影响因子：
5.4
作者：
Staskus, KA;Zhong, WD;Hasse, AT
通讯作者：
Hasse, AT